外消旋紫草素肟衍生物的设计合成及其抗肿瘤活性研究
发布时间：2018-08-03 12:04
【摘要】：紫草是我国的传统中药,有很长的临床应用历史。紫草中的主要药用成分是紫草素和其对映体阿卡宁及其酯类化合物。目前,紫草素虽然有着较强的抗肿瘤作用,但其做为抗肿瘤药物直接在临床上应用还有很长的路要走,这主要是因为这类化合物的选择性差,细胞毒作用太大,不仅抑制肿瘤细胞生长,也抑制大量正常细胞的生长。所以如何降低紫草素类化合物对正常细胞的毒性,提高对癌细胞的选择性将是开发临床上能应用的此类抗肿瘤药物亟待解决的问题。我们对紫草素进行结构改造,希望能找到一种更高效、对正常细胞低毒的抗肿瘤药物。设计一系列外消旋紫草肟衍生物。通过全合成得到中间体外1,4,5,8-氧-四甲基消旋紫草素,1,4,5,8-氧-四甲基消旋紫草素侧链羟基经过酯化或醚化,再经硝酸铈铵脱甲基氧化得到2位和6位二羰基氧二甲基侧链羟基酯化或醚化的紫草素,进一步通过萘环母核羰基成肟修饰后得到外消旋紫草素肟衍生物。我们共设计并合成了28个新化合物,包括紫草素1,4-二羰基双肟化合物21个,紫草素4-羰基单肟化合物7个,其中侧链成酯化合物22个,成醚化合物6个。我们通过MTT法测得其对MCF-7,K-562与DU-145人体肿瘤细胞的抑制活性。体外活性数据显示肟系列的紫草素衍生物对MCF-7,K-562两种肿瘤细胞抑制活性较好,而对DU-145肿瘤细胞抑制作用较差,显示出一定的作用选择性。5,8-氧二甲基紫草素的羰基成肟后,肿瘤抑制活性反而增强,活性顺序为:1,4-二羰基肟(双肟)4-羰基肟(单肟)二羰基化合物,肟衍生物的体外活性强于其前体醌。另外,成肟后,侧链对活性影响较大,侧链羟基成酯后活性明显提高,成醚效果较差。其中化合物2DMASKO-5活性比较好,对人乳腺癌细胞MCF-7和人白血病细胞K562的抑制IC50分别为12.7μM和1.3μM。我们选择该化合物进行了体内抗肿瘤作用研究。体内实验结果显示,2-DMSAKO-5腹腔给药12mg/kg对小鼠EMT6乳腺癌移植瘤抑瘤率为41.4%,对小鼠CT26结肠癌移植瘤抑瘤率为45.9%,活性均高于阳性对照组5-FU腹腔给药25 mg/kg组,而对小鼠S180组移植瘤抑瘤活性比较差,没有阳性对照组效果好。而且2-DMSAKO-5给药组小鼠体重增长与阴性对照组相同,未发现与紫草素及其二甲基衍生物体内试验所产生的局部刺激、腹水等类似毒性。通过对紫草素萘茜母核羟基甲基化、羰基肟化修饰,成功地解决了困扰紫草素作为抗肿瘤药物的毒性和选择性问题,通过进一步的侧链结构优化和作用机制的深入研究,有望得到活性、选择性及成药性更好的候选化合物进入临床前研究,为开发结构新颖、机制独特、高效低毒的有自主知识产权的原创性抗肿瘤新药奠定了基础。
[Abstract]:Purple herb is a traditional Chinese medicine in China, which has a long history of clinical application. The main medicinal components of Shikonin and its enantiomers, acanine and their esters. At present, although Shikonin has a strong anti-tumor effect, it still has a long way to go as an antitumor drug in clinical application. This is mainly due to the poor selectivity and cytotoxic effect of these compounds. It not only inhibits the growth of tumor cells, but also inhibits the growth of a large number of normal cells. Therefore, how to reduce the toxicity of Shikonin compounds to normal cells and improve the selectivity to cancer cells will be the urgent problems to be solved in the development of this kind of anti-tumor drugs. In order to find a more efficient and low toxic antitumor drug, we modified the structure of Shikonin. A series of racemic oxime derivatives were designed. The side chain hydroxyl of the intermediate was esterified or etherified by the synthesis of the intermediates (1: 4, 4, 5, 5)-oxy-tetramethyl racemyl luciferin, and the side chain hydroxyl of the intermediate was esterified or etherified, and the hydroxyl of the side chain of the intermediate was esterified or etherized. The alkaloids of 2 and 6 position dicarbonyl oxomethyl side chain hydroxy esterification or etherification were obtained by cerium ammonium nitrate demethylation and then modified by naphthalene ring mother nucleus carbonyl oxime to obtain racemic porphyrin oxime derivatives. We have designed and synthesized 28 new compounds, including 21 porphyrin 4-dicarbonyl dioxime compounds, 7 porphyrin 4-carbonyl monooxime compounds, including 22 side chain ester forming compounds and 6 ether forming compounds. The inhibitory activity of MCF-7 K-562 and DU-145 human tumor cells was determined by MTT assay. The in vitro activity data showed that the oxime derivatives had better inhibitory activity on MCF-7 and K-562 tumor cells than on DU-145 tumor cells. The inhibitory activity of tumor was increased in the order of 1: 1, 4-dicarbonyl oxime (dioxime) 4-carbonyl oxime (monoxime) dicarbonyl compound, and the activity of oxime derivative was stronger than that of its precursor in vitro. In addition, after oxime formation, the side chain has a great effect on the activity, the side chain hydroxyl ester formation activity is obviously increased, and the ether formation effect is poor. The inhibitory IC50 of compound 2DMASKO-5 on MCF-7 and K562 cells were 12.7 渭 M and 1.3 渭 M, respectively. We selected the compound to study its anti-tumor effect in vivo. The results of in vivo experiment showed that the inhibitory rate of 12mg/kg on EMT6 breast cancer transplanted tumor was 41.4%, and on CT26 colon cancer transplantation tumor was 45.90.The activity was higher than that of the positive control group (25 mg/kg group). The tumor inhibitory activity of S 180 group was poor, and the effect of positive control group was better than that of positive control group. The weight gain of mice in 2-DMSAKO-5 group was the same as that in negative control group, and no similar toxicity was found with the local stimulation and ascites produced by the in vivo test of Shikonin and its dimethyl derivatives. The toxicity and selectivity of porphyrin as an antitumor drug were solved successfully by hydroxyl methylation and carbonyl oximation modification of the mother nucleus of porphyrin, and the side chain structure optimization and the mechanism of action were further studied. It is expected that candidate compounds with better activity, selectivity and drug formation will be used in preclinical research, which will lay a foundation for the development of novel antitumor drugs with unique structure, unique mechanism and high efficiency and low toxicity, with their own intellectual property rights.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R284.3;R285.5
本文编号：2161689
[Abstract]:Purple herb is a traditional Chinese medicine in China, which has a long history of clinical application. The main medicinal components of Shikonin and its enantiomers, acanine and their esters. At present, although Shikonin has a strong anti-tumor effect, it still has a long way to go as an antitumor drug in clinical application. This is mainly due to the poor selectivity and cytotoxic effect of these compounds. It not only inhibits the growth of tumor cells, but also inhibits the growth of a large number of normal cells. Therefore, how to reduce the toxicity of Shikonin compounds to normal cells and improve the selectivity to cancer cells will be the urgent problems to be solved in the development of this kind of anti-tumor drugs. In order to find a more efficient and low toxic antitumor drug, we modified the structure of Shikonin. A series of racemic oxime derivatives were designed. The side chain hydroxyl of the intermediate was esterified or etherified by the synthesis of the intermediates (1: 4, 4, 5, 5)-oxy-tetramethyl racemyl luciferin, and the side chain hydroxyl of the intermediate was esterified or etherified, and the hydroxyl of the side chain of the intermediate was esterified or etherized. The alkaloids of 2 and 6 position dicarbonyl oxomethyl side chain hydroxy esterification or etherification were obtained by cerium ammonium nitrate demethylation and then modified by naphthalene ring mother nucleus carbonyl oxime to obtain racemic porphyrin oxime derivatives. We have designed and synthesized 28 new compounds, including 21 porphyrin 4-dicarbonyl dioxime compounds, 7 porphyrin 4-carbonyl monooxime compounds, including 22 side chain ester forming compounds and 6 ether forming compounds. The inhibitory activity of MCF-7 K-562 and DU-145 human tumor cells was determined by MTT assay. The in vitro activity data showed that the oxime derivatives had better inhibitory activity on MCF-7 and K-562 tumor cells than on DU-145 tumor cells. The inhibitory activity of tumor was increased in the order of 1: 1, 4-dicarbonyl oxime (dioxime) 4-carbonyl oxime (monoxime) dicarbonyl compound, and the activity of oxime derivative was stronger than that of its precursor in vitro. In addition, after oxime formation, the side chain has a great effect on the activity, the side chain hydroxyl ester formation activity is obviously increased, and the ether formation effect is poor. The inhibitory IC50 of compound 2DMASKO-5 on MCF-7 and K562 cells were 12.7 渭 M and 1.3 渭 M, respectively. We selected the compound to study its anti-tumor effect in vivo. The results of in vivo experiment showed that the inhibitory rate of 12mg/kg on EMT6 breast cancer transplanted tumor was 41.4%, and on CT26 colon cancer transplantation tumor was 45.90.The activity was higher than that of the positive control group (25 mg/kg group). The tumor inhibitory activity of S 180 group was poor, and the effect of positive control group was better than that of positive control group. The weight gain of mice in 2-DMSAKO-5 group was the same as that in negative control group, and no similar toxicity was found with the local stimulation and ascites produced by the in vivo test of Shikonin and its dimethyl derivatives. The toxicity and selectivity of porphyrin as an antitumor drug were solved successfully by hydroxyl methylation and carbonyl oximation modification of the mother nucleus of porphyrin, and the side chain structure optimization and the mechanism of action were further studied. It is expected that candidate compounds with better activity, selectivity and drug formation will be used in preclinical research, which will lay a foundation for the development of novel antitumor drugs with unique structure, unique mechanism and high efficiency and low toxicity, with their own intellectual property rights.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R284.3;R285.5
【参考文献】
相关期刊论文 前1条
1 庞志功,汪宝琪,李海峰;β-紫草素复方的药代动力学研究[J];西安医科大学学报;2001年04期
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